Squash hybrid ex 13056682 and parents thereof

ABSTRACT

The invention provides seed and plants of squash hybrid EX 13056682 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of squash hybrid EX 13056682 and the parent lines thereof, and to methods for producing a squash plant produced by crossing such plants with themselves or with another squash plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants, including the fruit and gametes of such plants.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority of U.S. Provisional Appl. Ser. No. 61/572,301, filed Aug. 30, 2011, the entire disclosure of which is incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, more specifically, to the development of squash hybrid EX 13056682 and the inbred squash lines LEB47-5020 and ZGN-130-1028.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits in a single variety/hybrid. Such desirable traits may include any trait deemed beneficial by a grower and/or consumer, including greater yield, resistance to insects or disease, tolerance to environmental stress, and nutritional value.

Breeding techniques take advantage of a plant's method of pollination. There are two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred to the same or another flower of the same plant or plant variety. A plant cross-pollinates if pollen comes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny, a homozygous plant. A cross between two such homozygous plants of different genotypes produces a uniform population of hybrid plants that are heterozygous for many gene loci. Conversely, a cross of two plants each heterozygous at a number of loci produces a population of hybrid plants that differ genetically and are not uniform. The resulting non-uniformity makes performance unpredictable.

The development of uniform varieties requires the development of homozygous inbred plants, the crossing of these inbred plants, and the evaluation of the crosses. Pedigree breeding and recurrent selection are examples of breeding methods that have been used to develop inbred plants from breeding populations. Those breeding methods combine the genetic backgrounds from two or more plants or various other broad-based sources into breeding pools from which new lines and hybrids derived therefrom are developed by selfing and selection of desired phenotypes. The new lines and hybrids are evaluated to determine which of those have commercial potential.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a squash plant of the hybrid designated EX 13056682, the squash line LEB47-5020 or squash line ZGN-130-1028. Also provided are squash plants having all the physiological and morphological characteristics of such a plant. Parts of these squash plants are also provided, for example, including pollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.

In another aspect of the invention, a plant of squash hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 comprising an added heritable trait is provided. The heritable trait may comprise a genetic locus that is, for example, a dominant or recessive allele. In one embodiment of the invention, a plant of squash hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 is defined as comprising a single locus conversion. In specific embodiments of the invention, an added genetic locus confers one or more traits such as, for example, herbicide tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism. In further embodiments, the trait may be conferred by a naturally occurring gene introduced into the genome of a line by backcrossing, a natural or induced mutation, or a transgene introduced through genetic transformation techniques into the plant or a progenitor of any previous generation thereof. When introduced through transformation, a genetic locus may comprise one or more genes integrated at a single chromosomal location.

The invention also concerns the seed of squash hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028. The squash seed of the invention may be provided as an essentially homogeneous population of squash seed of squash hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028. Essentially homogeneous populations of seed are generally free from substantial numbers of other seed. Therefore, seed of hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 may be defined as forming at least about 97% of the total seed, including at least about 98%, 99% or more of the seed. The seed population may be separately grown to provide an essentially homogeneous population of squash plants designated EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028.

In yet another aspect of the invention, a tissue culture of regenerable cells of a squash plant of hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 is provided. The tissue culture will preferably be capable of regenerating squash plants capable of expressing all of the physiological and morphological characteristics of the starting plant, and of regenerating plants having substantially the same genotype as the starting plant. Examples of some of the physiological and morphological characteristics of the hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 include those traits set forth in the tables herein. The regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistils, flowers, seed and stalks. Still further, the present invention provides squash plants regenerated from a tissue culture of the invention, the plants having all the physiological and morphological characteristics of hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028.

In still yet another aspect of the invention, processes are provided for producing squash seeds, plants and fruit, which processes generally comprise crossing a first parent squash plant with a second parent squash plant, wherein at least one of the first or second parent squash plants is a plant of squash line LEB47-5020 or squash line ZGN-130-1028. These processes may be further exemplified as processes for preparing hybrid squash seed or plants, wherein a first squash plant is crossed with a second squash plant of a different, distinct genotype to provide a hybrid that has, as one of its parents, a plant of squash line LEB47-5020 or squash line ZGN-130-1028. In these processes, crossing will result in the production of seed. The seed production occurs regardless of whether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing” comprises planting seeds of a first and second parent squash plant, often in proximity so that pollination will occur for example, mediated by insect vectors. Alternatively, pollen can be transferred manually. Where the plant is self-pollinated, pollination may occur without the need for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first and second parent squash plants into plants that bear flowers. A third step may comprise preventing self-pollination of the plants, such as by emasculating the flowers (i.e., killing or removing the pollen).

A fourth step for a hybrid cross may comprise cross-pollination between the first and second parent squash plants. Yet another step comprises harvesting the seeds from at least one of the parent squash plants. The harvested seed can be grown to produce a squash plant or hybrid squash plant.

The present invention also provides the squash seeds and plants produced by a process that comprises crossing a first parent squash plant with a second parent squash plant, wherein at least one of the first or second parent squash plants is a plant of squash hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028. In one embodiment of the invention, squash seed and plants produced by the process are first generation (F₁) hybrid squash seed and plants produced by crossing a plant in accordance with the invention with another, distinct plant. The present invention further contemplates plant parts of such an F₁ hybrid squash plant, and methods of use thereof. Therefore, certain exemplary embodiments of the invention provide an F₁ hybrid squash plant and seed thereof.

In still yet another aspect, the present invention provides a method of producing a plant derived from hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028, the method comprising the steps of: (a) preparing a progeny plant derived from hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028, wherein said preparing comprises crossing a plant of the hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 with a second plant; and (b) crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation. In further embodiments, the method may additionally comprise: (c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a subsequent generation and crossing the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps for an additional 3-10 generations to produce a plant derived from hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028. The plant derived from hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 may be an inbred line, and the aforementioned repeated crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line. In the method, it may be desirable to select particular plants resulting from step (c) for continued crossing according to steps (b) and (c). By selecting plants having one or more desirable traits, a plant derived from hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 is obtained which possesses some of the desirable traits of the line/hybrid as well as potentially other selected traits.

In certain embodiments, the present invention provides a method of producing food or feed comprising: (a) obtaining a plant of squash hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028, wherein the plant has been cultivated to maturity, and (b) collecting at least one squash from the plant.

In still yet another aspect of the invention, the genetic complement of squash hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 is provided. The phrase “genetic complement” is used to refer to the aggregate of nucleotide sequences, the expression of which sequences defines the phenotype of, in the present case, a squash plant, or a cell or tissue of that plant. A genetic complement thus represents the genetic makeup of a cell, tissue or plant, and a hybrid genetic complement represents the genetic make up of a hybrid cell, tissue or plant. The invention thus provides squash plant cells that have a genetic complement in accordance with the squash plant cells disclosed herein, and seeds and plants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles, and by the expression of phenotypic traits that are characteristic of the expression of the genetic complement, e.g., isozyme typing profiles. It is understood that hybrid EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 could be identified by any of the many well known techniques such as, for example, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).

In still yet another aspect, the present invention provides hybrid genetic complements, as represented by squash plant cells, tissues, plants, and seeds, formed by the combination of a haploid genetic complement of a squash plant of the invention with a haploid genetic complement of a second squash plant, preferably, another, distinct squash plant. In another aspect, the present invention provides a squash plant regenerated from a tissue culture that comprises a hybrid genetic complement of this invention.

Any embodiment discussed herein with respect to one aspect of the invention applies to other aspects of the invention as well, unless specifically noted.

The term “about” is used to indicate that a value includes the standard deviation of the mean for the device or method being employed to determine the value. The use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive. When used in conjunction with the word “comprising” or other open language in the claims, the words “a” and “an” denote “one or more,” unless specifically noted otherwise. The terms “comprise,” “have” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “having,” “includes” and “including,” are also open-ended. For example, any method that “comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps. Similarly, any plant that “comprises,” “has” or “includes” one or more traits is not limited to possessing only those one or more traits and covers other unlisted traits.

Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and any specific examples provided, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants, seeds and derivatives of squash hybrid EX 13056682, squash line LEB47-5020 and squash line ZGN-130-1028. The designation LEB47-5020 as used herein has the same meaning as, and is used interchangeably with, the designation LEB-47-5020.

Squash hybrid EX 13056682 is a Long Lebanese type squash in the Eskenderany class with multiple virus resistance such as to Zucchini Yellow Mosaic Virus (ZYMV), Watermelon Mosaic Virus (WMV), and Squash leaf curl virus (SLCV). Hybrid EX 13056682 has high yield potential and exceptional fruit color conforming to market preferences.

ZGN-130-1028 is a green zucchini breeding line with exceptional levels of virus resistance. This genotype in particular shows high level resistance to geminiviruses, such as squash leaf curl virus (SLCV), as well as resistance to potyviruses, such as zucchini yellow mosaic virus (ZYMV), watermelon mosaic virus (WMV), and papaya ringspot virus (PRSV). In addition to the disease resistance, this genotype has a dark green fruit color preferred by growers in many regions, as well as acceptable horticultural characteristics.

A. Origin and Breeding History of Squash Hybrid EX 13056682

The parents of hybrid EX 13056682 are LEB47-5020 and ZGN-130-1028 (U.S. Patent Publication No. US 2008-0313755 A1). These parents were created as follows:

LEB47-5020 was developed through a minimum of 18 generations of inbreeding (controlled self pollination) and crossing. The initial source of the key traits of value (resistance to Zucchini Yellow Mosaic Virus (ZYMV) and Watermelon Mosaic Virus II (WMV-II)) was a diverse early generation population of squash distributed by Henry Munger of Cornell University in 1986 to collaborating seed companies and initially designated as “HMZYR” (presumably representing Henry Munger ZYMV resistant family). An individual from that population was crossed to a selection from the landrace “Small Green Algerian” in 1986 (generation 1), and then self pollinated for 3 generations, with selection for fruit color and shape in each generation (generations 2-4). An individual was selected from this population after the 3 generations of inbreeding and crossed to a proprietary breeding line known as LEB 47-98 (derived from a selection from cultivated landraces of squash common throughout the Middle East region). This germplasm was selected and self- and/or sib-pollinated for seven generations (generations 5-11), when an individual plant was crossed again to LEB-47-98 (generation 12). The progeny of this cross was selfed for 6 generations (12-17), when three individual plants were selected, self pollinated, bulked, and the bulk was identified as LEB47-5020. In generation 18 following the original selections this line was verified as phenotypically uniform for the purposes of commercial hybrid seed production.

The parent lines are uniform and stable, as is a hybrid produced therefrom. A small percentage of variants can occur within commercially acceptable limits for almost any characteristic during the course of repeated multiplication. However no variants are expected.

Squash line ZGN-130-1028 was developed by a selection strategy involving four components: (1) ZGN 130-1003, a breeding line contributing fruit quality characteristics and some potyvirus resistance, (2) ZGY 46-2604, the original gray zucchini source of resistance to SLCV, (3) G710, a breeding line contributing fruit quality characteristics, and (4) HP13HMW/HP134*1, a breeding line contributing high resistance to ZYMV, WMV-2, and PRSV.

ZGY 46-2604 was crossed to a series of breeding lines, including G710 in order to develop breeding populations with a combination of SLCV resistance and acceptable fruit types. The resulting F1 progeny of this cross was backcrossed to G710, and the BC1F2, BC1F3, and BC1F4 generations were selected for fruit and plant characteristics without regard to SLCV virus resistance in Tifton, Ga.

The self-pollinated BC1F5 progeny selections were included in a field trial in the Rio Grande Valley of Texas in late summer 2000 (a location and date chosen for its history of SLCV epidemics). Heavy SLCV pressure and high levels of potyvirus infestations in the same field allowed for the selections of breeding lines with SLCV and potyvirus resistance. Nineteen selections (or remnant seed in the case of failed pollinations) were chosen to be planted in the greenhouse at Woodland, Calif. in February, Year 1 and inoculated with SLCV. Four BC1F5 and BC1F6 families were selected for a high level of resistance to SLCV and for fruit quality characteristics.

The selected families were crossed to a series of thirty-one breeding lines in the spring of Year 1. During the summer of Year 1, these breeding lines were evaluated for virus resistance through individual and cocktail screens of potyviruses and for fruit quality characteristics in Woodland, Calif. Ten hybrids were selected, and these ten hybrids were subsequently evaluated for resistance to SLCV in a nursery planted in Hermosillo, Mexico in the later part of Year 1. The hybrid (ZGN130-1003/(ZGY46-2604/G710*1)) was selected for development of new germplasm. This hybrid was planted in the greenhouse at Woodland, Calif. in February of Year 2, and was crossed to line HP13HMW/HP134*1.

The resulting F1 population, known as (ZGN130-1003/(ZGY46-2604/G710*1F6))/(HP13HMW/HP134*1), was inoculated with a cocktail of ZYMV, WMV-2, and PRSV, and survivors were moved to the field in Woodland, Calif. in the summer of Year 2. This survivor population and all its progeny were named TAW. Thirty-seven individuals were selected from the TAW population based on potyvirus resistance and fruit and plant characteristics. The F2 generations from these thirty seven selections were sown in the greenhouse in Woodland, Calif. in January Year 3, and inoculated with ZYMV, WMV-2, PRSV, and SLCV.

The high SLCV resistance described was first observed in this early Year 3 screen, where F2 individuals in certain families showed much higher levels of resistance to SLCV than the original source of resistance, ZGY 46-2604. These F2 individuals were self-pollinated.

F3 families derived from the F2 individuals with unexpectedly high levels of SLCV resistance were inoculated with a cocktail of ZYMV, WMV-2, and PRSV, and resistant individuals were transplanted to a field near Woodland, Calif. in the summer of Year 3. One hundred and two F3 individuals were selected for self-pollination from this field based on their level of potyvirus resistance, and fruit and plant characteristics. Successful pollinations from the California F3 generation were sown in Florida to evaluate fruit quality and Sinaloa (for a simultaneous observation under heavy potyvirus and geminivirus natural pressure), and ten F4 families were selected. The field trial in Sinaloa allowed verification of the high level geminivirus resistance in an environment very similar to a commercial field. The selected lines provided a verification of the unexpected results from the early Year 3 greenhouse screen, because the F4 generation also showed a higher level of resistance to natural geminivirus infection than the genotype which was the original source of resistance.

Not all of the self-pollinations were successful in the F4 generation, thus some were recovered using remnant F4 seed in the first Woodland greenhouse cycle in January, Year 4. Both F4 and F5 families were included, as available, in this early Year 4 experiment, where selections were made following inoculation with SLCV (Table 1). These breeding lines also demonstrated a higher level of resistance to geminivirus than the original source of resistance. Self pollination of these selections from the first greenhouse cycle of Year 4 provided the F5 and F6 generation seeds used in the present experiment. The eight elite breeding families described in Table 1 show uniform, high resistance to SLCV. Each of the entries have the pedigree TAW. These entries represent 4 separate lineages from the F1 generation. The results found in some F4 and F5 selections from the TAW population were consistent with the observations in the F2 generation, and displayed the unique and valuable characteristics of these genotypes.

TABLE 1 Reaction to SLCV Inoculation in Year 4 Greenhouse Screen Pedigree Gen 1 2 3 4 5 AVG (TAW)-19-2-3-2 F5 13 1.0 (TAW)-19-2-3-3 F5 10 7 1.4 (TAW)-20-9-4-1 F5 4 3 1.4 (TAW)-4-1-1 F4 6 2 1.3 (TAW)-4-1-5 F4 1 2 1.7 (TAW)-15-1-7 F4 1 3 1.8 (TAW)-15-1-8 F4 2 5 1 1.9 (TAW)-30-6-7 F4 4 5 1.6 Grey Zucchini ADV 1 3 2 8 58 4.7 ZGY46-2604 ADV 3 11 24 20 1 3.1

In Table 1, the Pedigree column identifies the genotype tested. Grey Zucchini is widely understood to be a genotype fully susceptible to all viruses that infect Cucurbita pepo, and represents a positive control. The “Gen” gives the number of generations of self pollination, with “ADV” designating a uniform line of advanced generation. The columns labeled 1-5 indicate the severity of disease symptoms in an individual plant. The “Avg” column provides the average disease score for the genotype. The symptoms were evaluated on a 1 to 5 scale, with a value of 5 assigned to the most severe reaction following infection, corresponding to where newly developing leaves are curled, with a bright yellow mosaic pattern. Yellow patterns on leaves are concentrated around leaf margins in some genotypes. Successive leaves are similar, but smaller, leading to a rosette type habit around the growing tip(s) of the plant. Stunting develops rapidly, and will usually lead to necrosis of all growing tips. Infected plants sometimes produce a profusion of flowers, many of which do not open or do not open normally, often with highly reduced petals. The plant dies before fruiting. A SLCV score of 4 corresponds to the youngest leaves showing many spots and a curled-under form. Older leaves show severe mottling and distortion, but the plant may survive to produce some poorly developed and misshapen fruit. The SLCV score of 3 corresponds to the youngest leaf showing a small spot or two and is often curled under and the older leaves having obvious mottling and distortion, with a diffuse yellow mosaic pattern. Successive leaves beyond the first which shows symptoms show decreasing symptoms and in the weeks following the initial symptoms, normal growth habit returns to the plant. New leaves, beyond approximately the first five after initial symptom development, show very mild mosaic, but no curling. Late in the life cycle of the plant, when pollinated fruit are approaching physiological maturity, new leaves may once again show intermediate mosaic and mild curling. Reduction of petals on flowers may be exhibited, but this symptom is also transient. A SLCV score of 2 corresponds to some spotting appearing on the older leaves (those first showing symptoms described above) but not on the youngest leaves, as described for a SLCV resistant score of 3. Rather than five or more nodes before the disappearance of symptoms, reaction symptoms disappear between the second and fourth new leaf following symptom display. Fruit production is acceptable, with only mild discoloration. An SLCV score of 1 indicates that following infection a single newly developing leaf may show mild mosaic, but no curling. Successive leaves show no apparent symptoms of virus infection. Reduction of petals is not observed. Fruit yield of the plants is generally unaffected.

The F5 generation of the lineage leading to ZGN-130-1028 was self pollinated in the greenhouse from survivors of the SLCV screen described in Table 1. One additional generation of self pollination was performed on this lineage in Florida in late Year 4, so that the line ZGN-130-1028 represents all progeny of a single plant selection from the F6 generation, with pedigree (TAW)-19-2-3-2-1-6.

ZGN-130-1028 has a complement of resistance traits and horticultural characteristics that make it useful for many purposes, including for growing green zucchini hybrids adapted to Sonora and Sinaloa in Mexico, and also for growing long Lebanese type hybrids (similar to Eskenderany or Top Kapi), such as those used in North Africa and the Middle East.

The parent lines are uniform and stable, as is a hybrid produced therefrom. A small percentage of variants can occur within commercially acceptable limits for almost any characteristic during the course of repeated multiplication. However no variants are expected.

B. Physiological and Morphological Characteristics of Squash Hybrid EX 13056682, Squash Line LEB47-5020 and Squash Line ZGN-130-1028

In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of squash hybrid EX 13056682 and the parent lines thereof. A description of the physiological and morphological characteristics of such plants is presented in Tables 2-4.

TABLE 2 Physiological and Morphological Characteristics of Hybrid EX 13056682 Comparison Variety CHARACTERISTIC EX 13056682 Anita 1. Species Pepo Pepo 2. Kind/Use squash squash 3. Type summer (vegetable summer marrow) 4. Cotyledon length 28.6 mm 51.9 mm width 20.65 mm 30.5 mm apex rounded rounded veining plainly visible plainly visible color medium green light green color (RHS Color Chart) 141A 137B Seedling shape of cotyledons elliptic (Cora, Tivoli) elliptic intensity of green color of medium (Cora) light cotyledons cross section of cotyledons concave straight 5. Mature Plant growth habit bush bush plant type prickly prickly 6. Main Stem cross-section shape round round average diameter at mid-point 36.15 mm 29.2 mm of 1^(st) internode average length 27.5 cm 31.6 cm average number of internodes 19.75 23.5 Stem color partly green and partly partly green and partly yellow (Autumn Gold) yellow intensity of green color medium (Cinderella) medium mottling present (Cora) present tendrils absent to rudimentary absent to rudimentary (Goldrush, Sylvana) Plant growth habit bush (Greyzini) bush branching absent (Goldi) absent bush varieties only: attitude of erect to semi-erect semi-erect petiole (excluding lower (Sardane) external leaves) 7. Leaves blade shape reniform reniform blade form deep lobed deep lobed margin denticulate denticulate margin edges frilled frilled average width 37.15 cm 36.8 cm average length 28.6 cm 29 cm leaf surface smooth blistered dorsal surface pubescence soft hairy soft hairy vental surface pubescence bristled soft hairy color dark green dark green color (RHS Color Chart) 147A 139A leaf blotching blotched with gray blotched with gray leaf blade: size medium (Ambassador) large leaf blade: incisions medium (Jackpot) medium leaf blade: intensity of green dark (Everest) dark color of upper surface leaf blade: silvery patches present (Civac) present leaf blade: relative area small (Aziz) medium covered by silvery patches average petiole length 32.6 cm 33.05 cm petiole length medium (Goldi) medium petiole: number of prickles many (White Bush medium Scallop) 8. Flower pistillate flower: diameter 17.05 cm 13.7 cm pistillate flower: ovary drum-like drum-like pistillate flower: average 1.35 cm 2 cm pedicel length pistillate flower: margin shape curved curved pistillate flower: margin edges frilled plain pistillate flower: average sepal 1.05 mm 1.5 mm width pistillate flower: average sepal 7.9 mm 6.5 mm length pistillate flower: color deep yellow deep yellow pistillate flower: color (RHS 17A 17A Color Chart) staminate flower: average 14.35 mm 16.3 mm sepal length staminate flower: average 1.75 mm 2.5 mm sepal width staminate flower: average 154.8 mm 147 mm pedicel length staminate flower: color deep yellow deep yellow female flower: ring at inner present (Aurore) present side of corolla female flower: color of ring at yellow and green green inner side of corolla (Pueble) female flower: intensity of medium (Samba, medium green color of ring at inner Senator) side of corolla (varieties with green ring at inner side of corolla) male flower: ring at inner side present (Goldi) present of corolla male flower: color of ring at yellow and green green inner side of corolla (Alice, Carmina, Green Gem, Ibis) male flower: intensity of medium (Verdi) medium green color of ring at inner side of corolla (varieties with green ring at inner side of corolla) staminate flower: color 17A 21A 9. Fruit market maturity: average 17.15 cm 12.4 cm length market maturity: average 4.55 cm 3.8 cm width - stem end market maturity: average 4.4 cm 4.6 cm width - blossom end market maturity: average 249.5 gm 208.4 gm weight market maturity: shape straightneck straightneck according to variety type market maturity: apex taper pointed taper pointed market maturity: base flattened flattened market maturity: ribs inconspicuous inconspicuous market maturity: rib furrow shallow shallow depth market maturity: rib furrow medium wide narrow width market maturity: fruit surface smooth smooth market maturity: warts none none market maturity: blossom scar raised acorn raised acorn button young fruit: ratio length/ small (Opal) small maximum diameter (zucchini type varieties) young fruit: general shape tapered elliptical (Top pear shaped (zucchini and rounded Kapi) zucchini type varieties) young fruit: main color of partly white and partly partly white and partly skin (excluding color of ribs green green or grooves) young fruit: intensity of green light (Arlika) light color of skin (excluding color of ribs or grooves; only varieties with green color of skin) general shape club shaped pear shaped length (zucchini type medium (Cora) short varieties) maximum diameter (zucchini medium (Opal) large type varieties) ratio length/maximum small (Jedida) medium diameter (zucchini type varieties) blossom end (zucchini and rounded rounded neck type varieties) grooves present absent depth of grooves shallow (Connecticut Field) ribs present present protrusion of ribs medium (Ibis, Opal) weak main color of skin (excluding green (Ambassador, green color of dots, patches, stripes Baby Bear) and bands) intensity of green color of light very light skin (excluding color of dots, patches, stripes and bands; varieties with green color or skin) stripes in grooves absent (Baby Bear, absent Jack Be Little) color of ribs compared to same (Grey Zucchini) same main color of skin (excluding color of dots, patches, stripes and bands) dots present (Gold Rush, present Table Queen) size of main dots medium (Grey large Zucchini) secondary green color absent (Grey Zucchini, between ribs (excluding dots) Small Sugar) warts on skin absent absent size of flower scar medium (Spidi) medium length of peduncle very short (Arlesa) long color of peduncle green (Ambassador) green intensity of green color of light (Bianchini) medium peduncle mottling of peduncle present (Elite) present type rounded zucchini patches, stripes or bands in absent (Ambassador, ripe stage Black Jack) ripe fruit: main color of skin cream (Bianchini, yellow (excluding color of mottles, Opal) patches, stripes and bands) ripe fruit: secondary color of cream cream skin (excluding color of mottles, patches, stripes and bands) ripe fruit: green hue (only absent (Jedida) absent white and cream) ripe fruit: color of flesh cream (Elite) yellow ripe fruit: lignified rind present (Elite, Little present Gem, Scallopini, Yellow Summer Crookneck) ripe fruit: structure of flesh fibrous (Vegetable fibrous Spaghetti) 10. Rind average thickness at medial 2.45 mm 3.1 mm toughness hard hard overall color pattern regular irregular main or ground color creamy-buff creamy-yellow main or ground color (RHS 10C 20C Color Chart) color of streaks (RHS Color 142A 16A Chart) pattern of streaks not specific not specific color of spots buff creamy-brown color of spots (RHS Color 159C 162C Chart) pattern of spots not specific not specific 11. Flesh average blossom end 12.75 mm 14.4 mm thickness average medial thickness 63.45 mm 78.9 mm average stem end thickness 20.1 mm 17.6 mm texture (fine, granular, lumpy fine stringy or stringy) texture (soft, firm or brittle) soft firm texture (dry, moist or juicy) juicy moist flavor insipid insipid quality inedible good color whitish-cream cream color (RHS Color Chart) 157C 155C 12. Seed Cavity average length 31.1 cm 28.1 cm average width 7.1 cm 10.1 cm location conforms to fruit shape conforms to fruit shape placental tissues abundant abundant center core prominent prominent 13. Fruit Stalks average length 1.75 cm 3.3 cm average diameter 3 cm 2.5 cm cross-section shape irregular irregular twisting not twisted not twisted tapering tapered not tapered straightness straight straight texture hard spongy furrows deep deep surface spiny spiny attachment end expanded not expanded detaches with difficulty easily color light green medium green color (RHS Color Chart) 143B 144B 14. Seeds average length 15.6 mm 16.3 mm average width 9.12 mm 9.15 mm average thickness 2.77 mm 2.5 mm face surface smooth smooth color cream cream color (RHS Color Chart) 161C 162C luster glossy dull margin straight straight margin edge rounded rounded separation from pulp difficult easy average grams per 100 seeds 14.87 gm 16.5 gm average number of seeds per 301.5 346.5 fruit seed coat normal normal size very large (Citrouille large de Touraine) shape broad elliptic (Baby broad elliptic Boo) hull present (Baby Bear, present Elite) appearance of hull fully developed (Elite) rudimentary color of hull cream (De Nice à Fruit cream Rond) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

TABLE 3 Physiological and Morphological Characteristics of Line LEB47-5020 Comparison Variety CHARACTERISTIC LEB47-5020 Anita 1. Species Pepo Pepo 2. Kind/Use squash squash 3. Type summer (vegetable summer marrow) 4. Cotyledon length 46.9 mm 51.9 mm width 31.4 mm 30.5 mm apex rounded rounded veining plainly visible plainly visible color medium green light green color (RHS Color Chart) 137C 137B Seedling shape of cotyledons elliptic (Cora, Tivoli) elliptic intensity of green color of medium (Cora) light cotyledons cross section of cotyledons concave straight 5. Mature Plant growth habit bush bush plant type glabrous prickly 6. Main Stem cross-section shape angled round average diameter at mid-point 22.75 mm 29.2 mm of 1^(st) internode average length 42.75 cm 31.6 cm average number of internodes 22 23.5 Stem color completely green partly green and partly (Becky) yellow intensity of green color light (Bianchini) medium mottling present (Cora) present tendrils well developed (Baby absent to rudimentary Bear, Greyzini) Plant growth habit bush (Greyzini) bush branching absent (Goldi) absent bush varieties only: attitude of semi-erect to horizontal semi-erect petiole (excluding lower (Goldi) external leaves) 7. Leaves blade shape reniform reniform blade form shallow lobed deep lobed margin dentate denticulate margin edges frilled frilled average width 36.15 cm 36.8 cm average length 25.8 cm 29 cm leaf surface blistered blistered dorsal surface pubescence glabrous soft hairy vental surface pubescence bristled soft hairy color dark green dark green color (RHS Color Chart) 137B 139A leaf blotching not blotched blotched with gray leaf blade: size large (Kriti) large leaf blade: incisions shallow (Everest) medium leaf blade: intensity of green dark (Everest) dark color of upper surface leaf blade: silvery patches absent (Black Forest, present Scallopini) average petiole length 27.4 cm 33.05 cm petiole length medium (Goldi) medium petiole: number of prickles few (Opaline) medium 8. Flower pistillate flower: diameter 15.4 cm 13.7 cm pistillate flower: ovary turbinate drum-like pistillate flower: average 1.75 cm 2 cm pedicel length pistillate flower: margin shape curved curved pistillate flower: margin edges frilled plain pistillate flower: average sepal 1.55 mm 1.5 mm width pistillate flower: average sepal 8.55 mm 6.5 mm length pistillate flower: color deep yellow deep yellow pistillate flower: color (RHS 17A 17A Color Chart) staminate flower: average 14.95 mm 16.3 mm sepal length staminate flower: average 2.8 mm 2.5 mm sepal width staminate flower: average 141.6 mm 147 mm pedicel length staminate flower: color deep yellow deep yellow female flower: ring at inner present (Aurore) present side of corolla female flower: color of ring at green (Aurore, Early green inner side of corolla White Bush Scallop, President) female flower: intensity of medium (Samba, medium green color of ring at inner Senator) side of corolla (varieties with green ring at inner side of corolla) male flower: ring at inner side present (Goldi) present of corolla male flower: color of ring at green (Austral, Belor, green inner side of corolla Goldi) male flower: intensity of medium (Verdi) medium green color of ring at inner side of corolla (varieties with green ring at inner side of corolla) staminate flower: color 17A 21A 9. Fruit market maturity: average 13.75 cm 12.4 cm length market maturity: average 4.6 cm 3.8 cm width - stem end market maturity: average 4.15 cm 4.6 cm width - blossom end market maturity: average 230 gm 208.4 gm weight market maturity: shape straightneck straightneck according to variety type market maturity: apex rounded taper pointed market maturity: base rounded flattened market maturity: ribs inconspicuous inconspicuous market maturity: rib furrow shallow shallow depth market maturity: rib furrow narrow narrow width market maturity: fruit surface smooth smooth market maturity: warts none none market maturity: blossom scar slightly extended raised acorn button young fruit: ratio length/ medium (Cora) small maximum diameter (zucchini type varieties) young fruit: general shape pear shaped (Clarita) pear shaped (zucchini and rounded zucchini type varieties) young fruit: main color of partly white and partly partly white and partly skin (excluding color of ribs green green or grooves) young fruit: intensity of green light (Arlika) light color of skin (excluding color of ribs or grooves; only varieties with green color of skin) general shape pear shaped pear shaped length (zucchini type long (Carlotta) short varieties) maximum diameter (zucchini medium (Opal) large type varieties) ratio length/maximum medium (Cora) medium diameter (zucchini type varieties) blossom end (zucchini and pointed rounded neck type varieties) grooves absent absent ribs absent present main color of skin (excluding green (Ambassador, green color of dots, patches, stripes Baby Bear) and bands) intensity of green color of very light very light skin (excluding color of dots, patches, stripes and bands; varieties with green color or skin) stripes in grooves absent (Baby Bear, absent Jack Be Little) color of ribs compared to same (Grey Zucchini) same main color of skin (excluding color of dots, patches, stripes and bands) dots present (Gold Rush, present Table Queen) size of main dots small (Ambassador) large secondary green color absent (Grey Zucchini, between ribs (excluding dots) Small Sugar) warts on skin absent absent size of flower scar medium (Spidi) medium length of peduncle long (Tivoli) long color of peduncle partly yellow and partly green green (Autumn Gold) intensity of green color of light (Bianchini) medium peduncle mottling of peduncle present (Elite) present type rounded zucchini patches, stripes or bands in absent (Ambassador, ripe stage Black Jack) ripe fruit: main color of skin green yellow (excluding color of mottles, patches, stripes and bands) ripe fruit: secondary color of green cream skin (excluding color of mottles, patches, stripes and bands) ripe fruit: color of flesh cream (Elite) yellow ripe fruit: lignified rind absent (Small Sugar, present Table Queen, Vegetable Spaghetti) ripe fruit: structure of flesh not fibrous (Elite) fibrous 10. Rind average thickness at medial 2.65 mm 3.1 mm toughness woody and tough hard overall color pattern regular irregular main or ground color buff-cream creamy-yellow main or ground color (RHS 158B 20C Color Chart) 11. Flesh average blossom end 13.35 mm 14.4 mm thickness average medial thickness 63.95 mm 78.9 mm average stem end thickness 18.4 mm 17.6 mm texture (fine, granular, lumpy fine stringy or stringy) texture (soft, firm or brittle) soft firm texture (dry, moist or juicy) juicy moist flavor insipid insipid quality good good color whitish-cream cream color (RHS Color Chart) 155D 155C 12. Seed Cavity average length 19.1 cm 28.1 cm average width 7.75 cm 10.1 cm location conforms to fruit shape conforms to fruit shape placental tissues abundant abundant center core inconspicuous prominent 13. Fruit Stalks average length 2.3 cm 3.3 cm average diameter 2 cm 2.5 cm cross-section shape irregular irregular twisting not twisted not twisted tapering not tapered not tapered straightness straight straight texture spongy spongy furrows deep deep surface rough spiny attachment end slightly expanded not expanded detaches easily easily color light green medium green color (RHS Color Chart) 144B 144B 14. Seeds average length 15.7 mm 16.3 mm average width 9.95 mm 9.15 mm average thickness 2.45 mm 2.5 mm face surface smooth smooth color cream cream color (RHS Color Chart) 162D 162C luster dull dull margin straight straight margin edge rounded rounded separation from pulp easy easy average grams per 100 seeds 14.35 gm 16.5 gm average number of seeds per 137.5 346.5 fruit seed coat normal normal size medium (Diamant) large shape broad elliptic (Baby broad elliptic Boo) hull present (Baby Bear, present Elite) appearance of hull fully developed (Elite) rudimentary color of hull cream (De Nice à Fruit cream Rond) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

TABLE 4 Physiological and Morphological Characteristics of Line ZGN-130-1028 Average seed weight 0.15 grams Internode length Short Leaf silvering Medium Leaf serration Medium Leaf lobing Heavy Mature fruit color Green Mature fruit speckling Absent Plant type Bush Plant habit Open Maturity Mid season Skin texture Smooth Fruit type Zucchini Immature fruit color Green Immature fruit speckling Fine, light Fruit shape Cylindrical Seed coat Present ZYMV resistance level High WMV resistance level High PRSV resistance level High SLCV resistance level High Cucumber Mosaic Virus (CMV) resistance level Low Powdery mildew (Sphaerotheca fuliginella) resistance Low Petiole length Long Blossom scar Medium Transgenes None *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

C. Breeding Squash Plants

One aspect of the current invention concerns methods for producing seed of squash hybrid EX 13056682 involving crossing squash lines LEB47-5020 and ZGN-130-1028. Alternatively, in other embodiments of the invention, hybrid EX 13056682, line LEB47-5020, or line ZGN-130-1028 may be crossed with itself or with any second plant. Such methods can be used for propagation of hybrid EX 13056682 and/or the squash lines LEB47-5020 and ZGN-130-1028, or can be used to produce plants that are derived from hybrid EX 13056682 and/or the squash lines LEB47-5020 and ZGN-130-1028. Plants derived from hybrid EX 13056682 and/or the squash lines LEB47-5020 and ZGN-130-1028 may be used, in certain embodiments, for the development of new squash varieties.

The development of new varieties using one or more starting varieties is well known in the art. In accordance with the invention, novel varieties may be created by crossing hybrid EX 13056682 followed by multiple generations of breeding according to such well known methods. New varieties may be created by crossing with any second plant. In selecting such a second plant to cross for the purpose of developing novel lines, it may be desired to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Once initial crosses have been made, inbreeding and selection take place to produce new varieties. For development of a uniform line, often five or more generations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way of double-haploids. This technique allows the creation of true breeding lines without the need for multiple generations of selfing and selection. In this manner true breeding lines can be produced in as little as one generation. Haploid embryos may be produced from microspores, pollen, anther cultures, or ovary cultures. The haploid embryos may then be doubled autonomously, or by chemical treatments (e.g. colchicine treatment). Alternatively, haploid embryos may be grown into haploid plants and treated to induce chromosome doubling. In either case, fertile homozygous plants are obtained. In accordance with the invention, any of such techniques may be used in connection with a plant of the invention and progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossing transfers a specific desirable trait from one inbred or non-inbred source to an inbred that lacks that trait. This can be accomplished, for example, by first crossing a superior inbred (A) (recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate locus or loci for the trait in question. The progeny of this cross are then mated back to the superior recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny have the characteristic being transferred, but are like the superior parent for most or almost all other loci. The last backcross generation would be selfed to give pure breeding progeny for the trait being transferred.

The plants of the present invention are particularly well suited for the development of new lines based on the elite nature of the genetic background of the plants. In selecting a second plant to cross with EX 13056682 and/or squash lines LEB47-5020 and ZGN-130-1028 for the purpose of developing novel squash lines, it will typically be preferred to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Examples of desirable traits may include, in specific embodiments, high seed yield, high seed germination, seedling vigor, high fruit yield, disease tolerance or resistance, and adaptability for soil and climate conditions. Consumer-driven traits, such as a fruit shape, color, texture, and taste are other examples of traits that may be incorporated into new lines of squash plants developed by this invention.

D. Performance Characteristics

As described above, hybrid EX 13056682 exhibits desirable traits, as conferred by squash lines LEB47-5020 and ZGN-130-1028. The performance characteristics of hybrid EX 13056682 and squash lines LEB47-5020 and ZGN-130-1028 were the subject of an objective analysis of the performance traits relative to other varieties. The results of the analysis are presented below.

TABLE 5 Objective and Subjective performance characteristics of commercial and pre-commercial long Lebanese type hybrids collected in De'ir Alla, Jordan in 2011. Sowing date: 28 Dec. 2010, evaluation 24 Feb.-6 Apr., 2011 Hybrid YFSHP IFCLR MFUNF BES FLATT HRVEZ PLTVG PLHAB SPINE FINAL Frt val/plant ESKENDERANY 5 6 2 2 3 2 3 3 6 5 8.96 ESKENDERANY 5 6 2 3 7 3 3 3 6 5 9.77 ESKENDERANY 6 5 3 3 6 3 4 2 4 6 7.64 SVR13056682 3 2 4 4 6 3 3 3 4 4 8.32 SVR13056682 5 4 5 4 5 4 2 3 5 4 8.78 SVR13056682 5 3 3 5 6 2 3 3 6 5 9.44 YFSHP—young fruit shape, scale of 1 to 9 IFCLR—uniformity and appropriateness of fruit color at marketable (immature) stage, 1-9 scale MFUNF—uniformity of harvested fruit shape at marketable (immature) stage, 1-9 scale BES—blossom end scar size, 1-9 scale FLATT—Flower attachment strength, 1-9 scale HRVEZ—Ease of harvest, 1-9 scale PLTVG—plant vigor, scale of 1 to 9 PLHAB—plant growth habit desirability, scale of 1 to 9 SPINE—size and density of spines on foliage, scale of 1 to 9 FINAL—overall subjective opinion of the marketability of the variety, scale of 1 to 9 Frt val/plant—number of marketable fruits produced per plant in this plot

E. Further Embodiments of the Invention

In certain aspects of the invention, plants described herein are provided modified to include at least a first desired heritable trait. Such plants may, in one embodiment, be developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to a genetic locus transferred into the plant via the backcrossing technique. The term single locus converted plant as used herein refers to those squash plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to the single locus transferred into the variety via the backcrossing technique. By essentially all of the morphological and physiological characteristics, it is meant that the characteristics of a plant are recovered that are otherwise present when compared in the same environment, other than an occasional variant trait that might arise during backcrossing or direct introduction of a transgene.

Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety. The parental squash plant which contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent. This terminology refers to the fact that the nonrecurrent parent is used one time in the backcross protocol and therefore does not recur. The parental squash plant to which the locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as it is used for several rounds in the backcrossing protocol.

In a typical backcross protocol, the original variety of interest (recurrent parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be transferred. The resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a squash plant is obtained wherein essentially all of the morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred locus from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for a successful backcrossing procedure. The goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original variety. To accomplish this, a single locus of the recurrent variety is modified or substituted with the desired locus from the nonrecurrent parent, while retaining essentially all of the rest of the desired genetic, and therefore the desired physiological and morphological constitution of the original variety. The choice of the particular nonrecurrent parent will depend on the purpose of the backcross; one of the major purposes is to add some commercially desirable trait to the plant. The exact backcrossing protocol will depend on the characteristic or trait being altered and the genetic distance between the recurrent and nonrecurrent parents. Although backcrossing methods are simplified when the characteristic being transferred is a dominant allele, a recessive allele, or an additive allele (between recessive and dominant), may also be transferred. In this instance it may be necessary to introduce a test of the progeny to determine if the desired characteristic has been successfully transferred.

In one embodiment, progeny squash plants of a backcross in which a plant described herein is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of squash the recurrent parent as determined at the 5% significance level when grown in the same environmental conditions.

New varieties can also be developed from more than two parents. The technique, known as modified backcrossing, uses different recurrent parents during the backcrossing. Modified backcrossing may be used to replace the original recurrent parent with a variety having certain more desirable characteristics or multiple parents may be used to obtain different desirable characteristics from each.

With the development of molecular markers associated with particular traits, it is possible to add additional traits into an established germ line, such as represented here, with the end result being substantially the same base germplasm with the addition of a new trait or traits. Molecular breeding, as described in Moose and Mumm, 2008 (Plant Physiology, 147: 969-977), for example, and elsewhere, provides a mechanism for integrating single or multiple traits or QTL into an elite line. This molecular breeding-facilitated movement of a trait or traits into an elite line may encompass incorporation of a particular genomic fragment associated with a particular trait of interest into the elite line by the mechanism of identification of the integrated genomic fragment with the use of flanking or associated marker assays. In the embodiment represented here, one, two, three or four genomic loci, for example, may be integrated into an elite line via this methodology. When this elite line containing the additional loci is further crossed with another parental elite line to produce hybrid offspring, it is possible to then incorporate at least eight separate additional loci into the hybrid. These additional loci may confer, for example, such traits as a disease resistance or a fruit quality trait. In one embodiment, each locus may confer a separate trait. In another embodiment, loci may need to be homozygous and exist in each parent line to confer a trait in the hybrid. In yet another embodiment, multiple loci may be combined to confer a single robust phenotype of a desired trait.

Many single locus traits have been identified that are not regularly selected for in the development of a new inbred but that can be improved by backcrossing techniques. Single locus traits may or may not be transgenic; examples of these traits include, but are not limited to, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect resistance, modified fatty acid or carbohydrate metabolism, and altered nutritional quality. These comprise genes generally inherited through the nucleus.

Direct selection may be applied where the single locus acts as a dominant trait. For this selection process, the progeny of the initial cross are assayed for viral resistance and/or the presence of the corresponding gene prior to the backcrossing. Selection eliminates any plants that do not have the desired gene and resistance trait, and only those plants that have the trait are used in the subsequent backcross. This process is then repeated for all additional backcross generations.

Selection of squash plants for breeding is not necessarily dependent on the phenotype of a plant and instead can be based on genetic investigations. For example, one can utilize a suitable genetic marker which is closely genetically linked to a trait of interest. One of these markers can be used to identify the presence or absence of a trait in the offspring of a particular cross, and can be used in selection of progeny for continued breeding. This technique is commonly referred to as marker assisted selection. Any other type of genetic marker or other assay which is able to identify the relative presence or absence of a trait of interest in a plant can also be useful for breeding purposes. Procedures for marker assisted selection are well known in the art. Such methods will be of particular utility in the case of recessive traits and variable phenotypes, or where conventional assays may be more expensive, time consuming or otherwise disadvantageous. Types of genetic markers which could be used in accordance with the invention include, but are not necessarily limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).

F. Plants Derived by Genetic Engineering

Many useful traits that can be introduced by backcrossing, as well as directly into a plant, are those which are introduced by genetic transformation techniques. Genetic transformation may therefore be used to insert a selected transgene into a plant of the invention or may, alternatively, be used for the preparation of transgenes which can be introduced by backcrossing. Methods for the transformation of plants that are well known to those of skill in the art and applicable to many crop species include, but are not limited to, electroporation, microprojectile bombardment, Agrobacterium-mediated transformation and direct DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ either friable tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may transform immature embryos or other organized tissue directly. In this technique, one would partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading enzymes (pectolyases) or mechanically wound tissues in a controlled manner.

An efficient method for delivering transforming DNA segments to plant cells is microprojectile bombardment. In this method, particles are coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are concentrated on filters or solid culture medium. Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plant cells by acceleration is the Biolistics Particle Delivery System, which can be used to propel particles coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a surface covered with target cells. The screen disperses the particles so that they are not delivered to the recipient cells in large aggregates. Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system for introducing gene loci into plant cells. An advantage of the technique is that DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations (Klee et al., Bio-Technology, 3(7):637-642, 1985). Moreover, recent technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate the construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes. Additionally, Agrobacterium containing both armed and disarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene locus transfer. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S. Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and combinations of these treatments (see, e.g., Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature, 312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986; Marcotte et al., Nature, 335:454, 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. (Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl. Genet., 107:462-469, 2003).

A number of promoters have utility for plant gene expression for any gene of interest including but not limited to selectable markers, scoreable markers, genes for pest tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest. Examples of constitutive promoters useful for plant gene expression include, but are not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985), including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591, 1990; Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemly duplicated version of the CaMV 35S promoter, the enhanced 35S promoter (P-e35S); 1 the nopaline synthase promoter (An et al., Plant Physiol., 88:547, 1988); the octopine synthase promoter (Fromm et al., Plant Cell, 1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem; the cauliflower mosaic virus 19S promoter; a sugarcane bacilliform virus promoter; a commelina yellow mottle virus promoter; and other plant DNA virus promoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can also be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., Plant Physiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter, Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones, such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4) wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5) chemicals such as methyl jasmonate, salicylic acid, or Safener. It may also be advantageous to employ organ-specific promoters (e.g., Roshal et al., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988; Bustos et al., Plant Cell, 1:839, 1989).

Exemplary nucleic acids which may be introduced to plants of this invention include, for example, DNA sequences or genes from another species, or even genes or sequences which originate with or are present in the same species, but are incorporated into recipient cells by genetic engineering methods rather than classical reproduction or breeding techniques. However, the term “exogenous” is also intended to refer to genes that are not normally present in the cell being transformed, or perhaps simply not present in the form, structure, etc., as found in the transforming DNA segment or gene, or genes which are normally present and that one desires to express in a manner that differs from the natural expression pattern, e.g., to over-express. Thus, the term “exogenous” gene or DNA is intended to refer to any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar gene may already be present in such a cell. The type of DNA included in the exogenous DNA can include DNA which is already present in the plant cell, DNA from another plant, DNA from a different organism, or a DNA generated externally, such as a DNA sequence containing an antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and could potentially be introduced into a squash plant according to the invention. Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a squash plant include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest tolerance such as genes for fungal disease control, herbicide tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s). For example, structural genes would include any gene that confers insect tolerance including but not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference in their entirety. In another embodiment, the structural gene can confer tolerance to the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes by encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for example, Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito, Mol. Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNA which expresses a phenotype or morphology change of interest is useful for the practice of the present invention.

G. Definitions

In the description and tables herein, a number of terms are used. In order to provide a clear and consistent understanding of the specification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all of which alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F₁), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes from different plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation of the organs with a cytoplasmic or nuclear genetic factor or a chemical agent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenic plants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.

Marker: A readily detectable phenotype, preferably inherited in codominant fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no environmental variance component, i.e., heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” are used interchangeably to describe plants that show no symptoms to a specified biotic pest, pathogen, abiotic influence or environmental condition. These terms are also used to describe plants showing some symptoms but that are still able to produce marketable product with an acceptable yield. Some plants that are referred to as resistant or tolerant are only so in the sense that they may still produce a crop, even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Royal Horticultural Society (RHS) color chart value: The RHS color chart is a standardized reference which allows accurate identification of any color. A color's designation on the chart describes its hue, brightness and saturation. A color is precisely named by the RHS color chart by identifying the group name, sheet number and letter, e.g., Yellow-Orange Group 19A or Red Group 41B.

Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a squash variety are recovered in addition to the characteristics of the single locus transferred into the variety via the backcrossing technique and/or by genetic transformation.

Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.

Tissue Culture: A composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant.

Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a squash plant by transformation.

H. Deposit Information

A deposit of squash hybrid EX 13056682 and inbred parent lines LEB47-5020 and ZGN-130-1028, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date of deposit were Sep. 23, 2011, Sep. 23, 2011, and Apr. 3, 2007. The accession numbers for those deposited seeds of squash hybrid EX 13056682 and inbred parent lines LEB47-5020 and ZGN-130-1028 are ATCC Accession No. PTA-12115, ATCC Accession No. PTA-12118, and ATCC Accession No. PTA-8396, respectively. Upon issuance of a patent, all restrictions upon the deposits will be removed, and the deposits are intended to meet all of the requirements of 37 C.F.R. §1.801-1.809. The deposits will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary during that period.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the invention, as limited only by the scope of the appended claims.

All references cited herein are hereby expressly incorporated herein by reference. 

1. A squash plant comprising at least a first set of the chromosomes of squash line LEB47-5020, a sample of seed of said line having been deposited under ATCC Accession No. PTA-12118.
 2. A seed comprising at least a first set of the chromosomes of squash line LEB47-5020, a sample of seed of said line having been deposited under ATCC Accession No. PTA-12118.
 3. The plant of claim 1, which is hybrid.
 4. The plant of claim 3, wherein the hybrid plant is squash hybrid EX 13056682, a sample of seed of said hybrid having been deposited under ATCC Accession No. PTA-12115.
 5. A plant part of the plant of claim
 1. 6. The plant part of claim 5, further defined as a leaf, a ovule, pollen, a fruit, or a cell.
 7. The plant part of claim 6, further defined as a fruit.
 8. A squash plant, or a part thereof, having all the physiological and morphological characteristics of the squash plant of claim
 1. 9. A squash plant, or a part thereof, having all the physiological and morphological characteristics of the squash plant of claim
 4. 10. A tissue culture of regenerable cells of the plant of claim
 1. 11. The tissue culture according to claim 10, comprising cells or protoplasts from a plant part selected from the group consisting of embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks.
 12. A squash plant regenerated from the tissue culture of claim
 11. 13. A method of vegetatively propagating the plant of claim 1 comprising the steps of: (a) obtaining tissue capable of being propagated from a plant according to claim 1; (b) cultivating said tissue to obtain proliferated shoots; and (c) rooting said proliferated shoots to obtain rooted plantlets.
 14. The method of claim 13, further comprising growing plants from said rooted plantlets.
 15. A method of introducing a desired trait into a squash line comprising: (a) crossing a plant of line LEB47-5020, a sample of seed of said line having been deposited under ATCC Accession No. PTA-12118, with a second squash plant that comprises a desired trait to produce F1 progeny; (b) selecting an F1 progeny that comprises the desired trait; (c) crossing the selected F1 progeny with a plant of line LEB47-5020 to produce backcross progeny; and (d) repeating steps (b) and (c) three or more times to produce selected fourth or higher backcross progeny that comprise the desired trait.
 16. A squash plant produced by the method of claim
 15. 17. A method of producing a plant comprising a transgene, the method comprising introducing a transgene into a plant of squash hybrid EX 13056682 or squash line LEB47-5020, a sample of seed of said hybrid and line having been deposited under ATCC Accession No. PTA-12115 and ATCC Accession No. PTA-12118, respectively.
 18. A plant of squash hybrid EX 13056682 or squash line LEB47-5020 further comprising a transgene, a sample of seed of said hybrid and line having been deposited under ATCC Accession No. PTA-12115 and ATCC Accession No. PTA-12118, respectively.
 19. A seed that produces the plant of claim
 18. 20. A plant of squash hybrid EX 13056682 or squash line LEB47-5020 comprising a single locus conversion, a sample of seed of said hybrid and line having been deposited under ATCC Accession No. PTA-12115 and ATCC Accession No. PTA-12118, respectively.
 21. A seed that produced the plant of claim
 20. 22. A method for producing a seed of a plant derived from hybrid EX 13056682 or line LEB47-5020 comprising the steps of: (a) crossing a squash plant of hybrid EX 13056682 or line LEB47-5020 with a second squash plant; a sample of seed of said hybrid and line having been deposited under ATCC Accession No. PTA-12115 and ATCC Accession No. PTA-12118, respectively; and (b) allowing seed of a hybrid EX 13056682 or line LEB47-5020-derived squash plant to form.
 23. The method of claim 22, further comprising the steps of: (c) crossing a plant grown from said hybrid EX 13056682 or LEB47-5020-derived squash seed with itself or a second squash plant to yield additional hybrid EX 13056682 or LEB47-5020-derived squash seed; (d) growing said additional hybrid EX 13056682 or LEB47-5020-derived squash seed of step (c) to yield additional hybrid EX 13056682 or LEB47-5020-derived squash plants; and (e) repeating the crossing and growing steps of (c) and (d) to generate at least a first further hybrid EX 13056682 or LEB47-5020-derived squash plant.
 24. The method of claim 22, wherein the second squash plant is of an inbred squash line.
 25. The method of claim 23, further comprising: (f) crossing the further hybrid EX 13056682 or LEB47-5020-derived squash plant with a second squash plant to produce seed of a hybrid progeny plant.
 26. A method of producing a squash comprising: (a) obtaining a plant according to claim 1, wherein the plant has been cultivated to maturity; and (b) collecting a squash from the plant.
 27. The method of claim 26, wherein the plant is a plant of squash hybrid EX 13056682, a sample of seed of said hybrid EX 13056682 having been deposited under ATCC Accession No. PTA-12115.
 28. A method of producing seed comprising crossing the plant of claim 1 with itself or a second plant. 